title: Zinc binding to the Tyr402 and His402 allotypes of complement factor H: possible implications for age-related macular degeneration.
abstract: The Tyr402His polymorphism of complement factor H (FH) with 20 short complement regulator (SCR) domains is associated with age-related macular degeneration (AMD). How FH contributes to disease pathology is not clear. Both FH and high concentrations of zinc are found in drusen deposits, the key feature of AMD. Heterozygous FH is inhibited by zinc, which causes FH to aggregate. Here, zinc binding to homozygous FH was studied. By analytical ultracentrifugation, large amounts of oligomers were observed with both the native Tyr402 and the AMD-risk His402 homozygous allotypes of FH and both the recombinant SCR-6/8 allotypes with Tyr/His402. X-ray scattering also showed that both FH and SCR-6/8 allotypes strongly aggregated at >10 μM zinc. The SCR-1/5 and SCR-16/20 fragments were less likely to bind zinc. These observations were supported by bioinformatics predictions. Starting from known zinc binding sites in crystal structures, we predicted 202 putative partial surface zinc binding sites in FH, most of which were in SCR-6. Metal site prediction web servers also suggested that SCR-6 and other domains bind zinc. Predicted SCR-6/8 dimer structures showed that zinc binding sites could be formed at the protein-protein interface that would lead to daisy-chained oligomers. It was concluded that zinc binds weakly to FH at multiple surface locations, most probably within the functionally important SCR-6/8 domains, and this explains why zinc inhibits FH activity. Given the high pathophysiological levels of bioavailable zinc present in subretinal deposits, we discuss how zinc binding to FH may contribute to deposit formation and inflammation associated with AMD.
authors.affiliation: Department of Structural and Molecular Biology, Division of Biosciences, Darwin Building, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Ruodan
authors.initials: R
authors.lastname: Nan
authors.forename: Irene
authors.initials: I
authors.lastname: Farabella
authors.forename: Felix F
authors.initials: FF
authors.lastname: Schumacher
authors.forename: Ami
authors.initials: A
authors.lastname: Miller
authors.forename: Jayesh
authors.initials: J
authors.lastname: Gor
authors.forename: Andrew C R
authors.initials: AC
authors.lastname: Martin
authors.forename: David T
authors.initials: DT
authors.lastname: Jones
authors.forename: Imre
authors.initials: I
authors.lastname: Lengyel
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Perkins
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D017242:Complement Factor H / Q000737:chemistry* / Q000235:genetics; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D008268:Macular Degeneration / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D015593:Retinal Drusen / Q000503:physiopathology; D012542:Scattering, Radiation; D014443:Tyrosine / Q000737:chemistry*; D014965:X-Rays; D015032:Zinc / Q000737:chemistry*
chemical_list: D014443:Tyrosine; D006639:Histidine; D017242:Complement Factor H; D015032:Zinc